title: Quantification of HBG mRNA in primary erythroid cultures: prediction of the response to hydroxyurea in sickle cell and beta-thalassemia.
abstract: OBJECTIVE
  Increased expression of fetal hemoglobin (HbF) may ameliorate the clinical course of hemoglobinopathies like sickle cell disease (SCD) and β-thalassemia. Hydroxyurea (HU) can stimulate HbF production in these diseases but the response is highly variable indicating the utility of developing an in vitro test to predict the patient's response to HU. We assessed whether the HbF response of patients with SCD and thalassemia intermedia (TI) to HU correlates with HBG (both γ-globin genes) expression in their cultured erythroid progenitors following exposure to HU.
  
  
  METHODS
  We exposed primary erythroid cultures from peripheral blood mononuclear cells from 30 patients with SCD and 15 with TI to HU and measured HBG mRNA by real-time quantitative PCR. The same patients were then treated with HU and their HbF response after treatment with a stable dose of HU was compared with the mRNA results in cultured cells.
  
  
  CONCLUSIONS
  The fold increase in HBG mRNA in erythroid progenitors was similar to the fold increase in HbF in vivo. Quantification of HBG mRNA in erythroid progenitor cell cultures from patients with SCD and TI is predictive of their clinical response to HU.
authors.affiliation: Dipartimento di Oncologia ed Ematologia, U.O.C. Ematologia per le Malattie Rare del Sangue e degli Organi Ematopoietici, A.O. Ospedali Riuniti Villa Sofia-Cervello, Palermo, Italy.
authors.forename: Alice
authors.initials: A
authors.lastname: Pecoraro
authors.forename: Paolo
authors.initials: P
authors.lastname: Rigano
authors.forename: Antonio
authors.initials: A
authors.lastname: Troia
authors.forename: Roberta
authors.initials: R
authors.lastname: Calzolari
authors.forename: Concetta
authors.initials: C
authors.lastname: Scazzone
authors.forename: Aurelio
authors.initials: A
authors.lastname: Maggio
authors.forename: Martin H
authors.initials: MH
authors.lastname: Steinberg
authors.forename: Rosalba
authors.initials: R
authors.lastname: Di Marzo
mesh_terms: D000328:Adult; D000368:Aged; D000755:Anemia, Sickle Cell / Q000097:blood / Q000188:drug therapy / Q000235:genetics*; D015672:Erythroid Precursor Cells / Q000378:metabolism*; D005260:Female; D005319:Fetal Hemoglobin / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D006918:Hydroxyurea / Q000008:administration & dosage / Q000627:therapeutic use; D008297:Male; D008875:Middle Aged; D061251:Primary Cell Culture; D012333:RNA, Messenger / Q000235:genetics; D016896:Treatment Outcome; D055815:Young Adult; D017086:beta-Thalassemia / Q000097:blood / Q000188:drug therapy / Q000235:genetics*; D055545:gamma-Globins / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D055545:gamma-Globins; D005319:Fetal Hemoglobin; D006918:Hydroxyurea
keywords: hydroxyurea; liquid erythroid cultures; real-time quantitative PCR; sickle cell disease; β-thalassemia